急性髓性白血病免疫球蛋白重链基因和T细胞受体γ基因重排的检测意义

免疫球蛋白重链基因(IgH)和T细胞受体(TCR)基因重排常被认为是淋巴细胞的克隆标志。但是，近年来的研究表明，急性髓性白血病(AML)细胞亦可出现IgH和TCR基因重排。为此，我们采集51例AML患者骨髓，利用PCR方法检测IgH及TCRγ基因重排，并初步探讨其临床意义。     

急性非淋巴细胞性白血病免疫球蛋白和T细胞受体基因重排的研究及其意义

采用PCR法对25例不同时期急性非淋巴细胞性白血病（ANLL）患者免疫球蛋白重链（IgH）和T细胞受体γ链（TcRr）基因重排进行了研究。结果显示，3例（12．0％）发生IgH基因重排，3例（12．0％）出现TcRr基因重排，其中1例同时出现IgH和TcRr基因重排，对这种ANLL中序列失真现象的机制及其在白血病基因分型及检测微小残留疾病中的意义进行讨论。     

急性淋巴细胞白血病细胞克隆性分析的临床意义

采用多聚酶链反应 (polymerase chain reaction,PCR)技术检测 49例急性淋巴细胞白血病患者免疫球蛋白重链 (imm unoglobulin heavy chain,Ig H)基因重排 ,并分析其细胞克隆性。结果 :35例显示有克隆性 Ig H重排 ,其中单克隆 2 6例 ,寡克隆 9例。9例完全缓解期中多次动态监测表明 :PCR检测 Ig H重排 ,在微小残留病监测和疾病复发判断中 ,具有重要的临床意义。     

免疫球蛋白重链基因重排检测在急性淋巴细胞白血病微小残留病中的应用

近30年来,随着化疗方案的改进,儿童急性淋巴细胞白血病(acute lymploblastic leukemia,ALL)的初治缓解率已达95%以上,但仍然有大约20%～30%的ALL缓解患儿复发。近十年研究表明,其复发的主要根源是体内仍残留有一定数量的白血病细胞,又称微小残留病变(minimal residual disease,     

急性非淋巴细胞性白血病免疫球蛋白和T细胞受体基因重排的研…

采用ＰＣＲ法对２５例不同时期急性非淋巴细胞性白血病患者免疫球蛋白重链和Ｔ细胞受体γ链基因重排进行了研究。结果显示，３例发生ＩｇＨ基因重排，３例出现ＴｃＲｒ基因重排，其中１例同时出现ＩｇＨ和ＴｃＲｒ基因重排，对这种ＡＮＬＬ中序列失真现象的机制及其在白血病基因分型豚检测微小残留疾病中的意义进行讨论。     

急性淋巴细胞白血病IgH超基因家族及T细胞受体δ、γ基因的研究

急性淋巴细胞白血病（ＡＬＬ）是一组血液系统的恶性克隆性疾病。目前,尽管应用强效化疗方案及骨髓移植等有效手段,使ＡＬＬ的疗效有较大改观。然而,诱导缓解后的复发问题仍是临床实践中的当务之急,而复发的根源是白血病克隆增殖的结果［１］。本研究以ＩｇＨ、Ｔ细胞受体（ＴＣＲ）γ和ＴＣＲδ基因重排为标志分子,用ＰＣＲ方法检测缓解后微小残留病（ＭＲＤ）,并探讨ＩｇＨ、ＴＣＲδ、γ基因重排在白血病免疫分型中的价值。 一、资料与方法 １．病例：３８例ＡＬＬ患者为初治的住院及门诊患者。全部患者均经临床、细胞形态学以及细…     

急性白血病受体基因重排的研究

目的：探讨急性白血病微小残留病（ＭＲＤ）与化疗效应差异及受体基因重排,在急性非淋巴细胞白血病（ＡＮＬＬ）中序列交叉现象及意义。方法：应用聚合酶链反应技术对免疫球蛋白（ＩｇＨ）及Ｔ细胞受体（ＴＣＲγ）基因重排进行定量测量。结果：４５例急性淋巴细胞白血病（ＡＬＬ）中阳性率８４．４％（３８／４５）,完全缓解（ＣＲ）后１８０ｄ检测,阳性率仍５５．６％（２５／４５）,２０例ＡＮＬＬ中５例检测到ＩｇＨ受体基因     

免疫分型结合基因重排对成人急性淋巴细胞白血病诊断及预后的判断价值

目的：探讨用免疫分型组合免疫球蛋白重链（IgH)及T细胞受体γ（TCRγ）基因重排对急性淋巴细胞白血病（ALL）的分型诊断及预后的判断价值。方法：免疫分型采用碱性磷酸酶抗性磷酸酶复合物（APAAP）免疫组化法,基因重排采用多聚酶链反应技术（PCR法）检测58例初治成人ALL患者。结果：①通过免疫分型检测,58例ALL中,43例（74．1％）为不带髓系相关标记的ALL（My^-ALL),15例（25．9％）为带髓系相关标记的ALL（My^ ALL),以CD15最常见。②采用PCR法检测IgH基因重排和TCRγ基因重排发现,58例ALL中有79．3％（46／58）免疫分型与基因重排结果完全吻合,即T－ALL出现TCRγ基因重排阳性,B－ALL出现IgH基因重排阳性,20．7％（12／58）基因重排结果与免疫分型不能完全吻合。③58例ALL经DOLP或DOCP方案1个疗程后,My^-ALL CR为72．1％（31／43）,My^ ALL为66．7％（10．15）;ALL不同阶段CR率分别是：T－ALL为82．4％（14／17）,ProB-ALL为50．0％（3／6）,C－ALL为90．5％（19／21）,RreB-ALL为33．3％（4／12）,成熟B－ALL为50．0％（1／2）;经基因重排检测与免疫分型吻合的ALL CR率为71．7％（33／46）,不吻合的ALL66．7％（8／12）。结论：对于白血病的分型应在FAB分型的基础上加用免疫分,可提高确诊率且对预后判断有价值;基因重排诊断仅有参考价值,对预后尚无指导意义。     

非霍奇金淋巴瘤患者外周血及骨髓中免疫球蛋白、T 细胞受体基因重排检测的意义

目的探讨非霍奇金淋巴瘤(NHL)患者骨髓及外周血中免疫球蛋白(Ig)、T细胞受体(TCR)基因单克隆重排检测的临床意义。方法以BIOMED-2引物系统及多重PCR方法对60例NHL患者骨髓或外周血标本进行Ig及TCR基因重排检测。结果 B细胞NHL中,39.02%检出Ig基因单克隆重排,T细胞NHL中36.84%检出TCR基因单克隆重排;14例早期与46例晚期患者的Ig和(或)TCR基因单克隆重排阳性率分别为28.57%和41.3%;11例低变恶性患者和49例中、高度恶性患者的Ig和(或)TCR基因单克隆重排阳性率分别为36.36%和38.78%;以上两者间比较差异均无统计学意义(P均>0.05)。53例NHL患者骨髓涂片检测骨髓侵犯阳性率为13.21%,明显低于骨髓基因重排检测阳性率(43.40%),两者比较差异有统计学意义(P<0.05)。结论骨髓及外周血Ig及TCR基因重排PCR检测有助于早期发现骨髓侵犯及MRD。     

三阴性骨髓纤维化伴免疫球蛋白重链基因重排一例

<正>患者,男,61岁,因“发现全血细胞减少9月余,乏力半月余,加重4天”于2020年4月18日入院。患者9个月前体检发现WBC、RBC、PLT计数均减少,其他无特殊不适,未予重视故未监测血常规。半月余前患者无明显诱因出现乏力,伴活动后气促,自觉尚可耐受,未就诊。4日前乏力及活动后气促较前明显加重,无咳嗽、发热、胸闷、胸痛、头晕、血尿、皮肤出血等,为求进一步治疗遂至我院,以“全血细胞减少查因”收入我科。既往史：两年前体检发现脂肪肝、脾脏肿大,     

不明原因发热患者检测淋巴细胞IgH和TCR基因重排的意义

目的:探讨不明原因发热(FUO)患者淋巴细胞免疫球蛋白重链(Ig H)和T细胞受体-γ(TCR-γ)基因重排的临床意义。方法:采用半巢式PCR方法检测110例FUO患者骨髓淋巴细胞Ig H和TCR-γ基因重排的阳性率。结果:84例淋巴瘤发热患者中,Ig H基因重排阳性34例,TCR-r基因重排阳性29例,两者阳性率达75%;26例非淋巴瘤发热患者中,Ig H基因重排阳性0例,TCR-r基因重排阳性1例,基因重排阳性率3.8%,差异有统计学意义(P0.05)。结论:对于FUO患者行基因重排有助于淋巴瘤的早期诊断,特别是对无法取得病理组织的患者意义更大。     

Crosslineage TCR delta rearrangements occur shortly after the DJ joinings of the IgH genes in childhood precursor B ALL and display age-specific characteristics

In order to test the hypothesis that the most immature T-cell receptor (TCR) rearrangements occur after the DJ joining of the immunoglobulin heavy chain genes (IgH), we analysed the TCR Vδ2–Dδ3 rearrangements in precursor B-cell leukaemias (PBC ALL) from 25 children younger than 3 years at disease onset and found that most of the junctional regions had N nucleotides inserted. We then selected 14 of these PCB ALLs for DJH (DJ joining of the IgH) characterization. These joining regions showed homology-directed recombination and lack of N regions, indicating absence of terminal deoxynucleotidyl transferase (TdT) activity during their rearrangement. Most leukaemias with a DJH rearrangement without N region have no, or only one, nucleotide in the joining regions of their Vδ2–Dδ3 rearrangements. The N regions of the TCR delta rearrangements displayed 'age-specific' differences: in children younger than 3 years of age the N regions were shorter than in those older than 3 years, and the rearrangements frequently contained complete segments. We conclude that the Vδ2–Dδ3 rearrangement in childhood PCB ALLs is an early event following DJH rearrangement and that it occurs shortly before or after the first hit, leading to malignant transformation.     

Various patterns of IgH deletion identified by FISH using combined IgH and IgH/CCND1 probes in multiple myeloma and chronic lymphocytic leukemia

Introduction: Interphase fluorescence in situ hybridization (FISH) can identify submicroscopic deletions adjacent to the breakpoints of rearrangements undetected by conventional cytogenetics. In this study, the characteristics and frequency of the IgH deletion identified by interphase FISH were investigated in patients with multiple myeloma (MM) and chronic lymphocytic leukemia (CLL). Methods: The study group included 29 patients with MM and eight patients with CLL. Interphase FISH was performed with the IgH dual color, break‐apart rearrangement probe and the IgH/CCND1 dual color, dual fusion translocation probe. Results: The IgH deletion was found in 14% (4/29) of patients with MM and 13% (1/8) of the patients with CLL. Four patients had deletions of the whole or variable region of IgH on the native chromosome 14, whereas one patient had a deletion of the IgH variable region on a der(11)t(11;14). In two patients, the IgH break‐apart FISH showed both patterns with and without IgH deletions. In cases showing the same pattern by IgH break‐apart FISH, the IgH/CCND1 FISH showed different patterns, and vice versa. Conclusion: A variety of patterns of the IgH deletion were identified by interphase FISH using IgH break‐apart and IgH/CCND1 probes in patients with MM and CLL. The results of this study suggest that the integrated information obtained with IgH break‐apart and IgH/CCND1 FISH was needed to interpret FISH results unambiguously.     

聚合酶链反应联合Southern杂交检测急性非淋巴细胞白血病IgH重排基因

免疫球蛋白重链（ＩｇＨ）重排，可作为Ｂ淋巴细胞白血病的基因标志，为进一步了解急性非淋巴细胞白血病（急非淋）ＩｇＨ重排情况，用聚合酶链反应（ＰＣＲ）联合地高辛标记ＪＨ探针Ｓｏｕｔｈｅｒｎ杂交，检测４１例急非淋病人ＩｇＨＣＤＲ－Ⅲ区域。７例（１７．１％）ＰＣＲ扩增阳性，此７例阳性病例均经Ｓｏｕｔｈｅｒｎ杂交证实，本实验的灵敏度为１０－４～１０－５水平。１２例完全缓解病例中３例（２５．０％）重排阳性，此３例均于半年内临床复发。结果表明，ＩｇＨ重排并不局限于Ｂ淋巴系白血病，也可发生于急非淋病人，其机制可能是，部分急非淋起源于较原始的多向造血祖细胞或除髓系白血病克隆外，尚存在亚克隆     

Maintenance of TCR clonality in T cells expressing genes for two TCR heterodimers

T cell receptor (TCR) allelic exclusion is believed to be primarily mediated by suppression of further recombination at the TCR locus after the expression of a functional TCR protein. Genetic allelic exclusion has been shown to be leaky for the beta chain and, more commonly, for the alpha chain. Here, we demonstrate an additional mechanism by which T cells can maintain monoclonality. T cells from double TCR transgenic mice express only one or the other of the two available TCRs at the cell surface. This "functional allelic exclusion" is apparently due to control of the TCR assembly process because these T cells express RNA and protein for all four transgenic TCR proteins. Lack of cell surface expression of the second TCR may be controlled by a failure to assemble the TCR heterodimer.     

Rapid and high-resolution detection of IgH gene rearrangements using PCR and melting curve analysis

The analytical methods of Southern blot hybridization (SBH) and the polymerase chain reaction (PCR) for complementarity determining region-3 (CDR3) are fundamental for detecting IgH gene rearrangement. However, there are problems stemming from the characteristics of both methods; especially, the long turn around time (TAT) because of the complex process in the SBH, and the low analytical sensitivity for amplicons in the PCR. Thus, to improve the PCR procedure, we investigated the application of detecting the clonal amplicons based on the different melting Temperature (T(m)) in internal melting domains corresponding to the CDR3 hypervariable region. Our new protocol is based on the combination of a LightCycler Technology with high-speed amplification, and Idaho-Technology with rapid and high-resolution melting curve analysis (MCA), designated PCR-MCA. This method can provide the results within 3 h with an analytical sensitivity of 10(-3). The diagnostic sensitivity and specificity relative to the results documented with the SBH analysis were 89.2% and 100%, respectively. This indicates that the new protocol of PCR-MCA is acceptable for clinical testing; especially, PCR-MCA is relevant in terms of the rapid and sensitive detection of IgH clonality within amplicons.